Azzindani/Open_Reaction_Data huggingface dataset:
Reaction SMILES: [CH3:1][O:2][C:3]1[CH:8]=[CH:7][C:6]([C:9]2[C:18]([C:19]3[CH:24]=[CH:23][C:22]([O:25][CH3:26])=[CH:21][CH:20]=3)=[N:17][C:16]3[C:11](=[CH:12][CH:13]=[C:14](/[CH:27]=[CH:28]/[C:29]([O:31]C)=[O:30])[CH:15]=3)[N:10]=2)=[CH:5][CH:4]=1.[OH-].[Na+]>CO.O>[CH3:1][O:2][C:3]1[CH:8]=[CH:7][C:6]([C:9]2[C:18]([C:19]3[CH:24]=[CH:23][C:22]([O:25][CH3:26])=[CH:21][CH:20]=3)=[N:17][C:16]3[C:11](=[CH:12][CH:13]=[C:14](/[CH:27]=[CH:28]/[C:29]([OH:31])=[O:30])[CH:15]=3)[N:10]=2)=[CH:5][CH:4]=1
Reactant: COC(=O)/C=C/c1ccc2nc(-c3ccc(OC)cc3)c(-c3ccc(OC)cc3)nc2c1
Agent: [Na+]
Agent: [OH-]
Solvent: O
Solvent: CO
Product: COc1ccc(-c2nc3ccc(/C=C/C(=O)O)cc3nc2-c2ccc(OC)cc2)cc1
Yield: 55.4 %

Procedure:
A solution of (E)-methyl 3-(2,3-bis(4-methoxyphenyl)quinoxalin-6-yl)acrylate (150 mg, 0.35 mmol, 1.00 equiv) and sodium hydroxide (70.4 mg, 1.76 mmol, 5.00 equiv) in methanol (10 mL) and H2O (2 ml) was stirred for 2 h at 50° C. in an oil bath. The resulting mixture was concentrated under vacuum, diluted with 20 mL of water. The pH value of the solution was adjusted to 3 with 1N hydrochloric acid. The solids were collected by filtration, and purified by re-crystallization from methanol, affording 80 mg (55%) of (E)-3-(2,3-bis(4-methoxyphenyl)quinoxalin-6-yl)acrylic acid as a yellow solid.
Reaction SMILES: Br[C:2]1[CH:3]=[N:4][C:5]([N:8]2[C:16]3[C:11](=[CH:12][CH:13]=[C:14]([C:17]([N:19]4[CH2:24][CH2:23][O:22][CH2:21][CH2:20]4)=[O:18])[CH:15]=3)[C:10]([CH2:25][OH:26])=[CH:9]2)=[N:6][CH:7]=1.[F:27][C:28]1[CH:33]=[CH:32][CH:31]=[CH:30][C:29]=1B(O)O>>[F:27][C:28]1[CH:33]=[CH:32][CH:31]=[CH:30][C:29]=1[C:2]1[CH:3]=[N:4][C:5]([N:8]2[C:16]3[C:11](=[CH:12][CH:13]=[C:14]([C:17]([N:19]4[CH2:24][CH2:23][O:22][CH2:21][CH2:20]4)=[O:18])[CH:15]=3)[C:10]([CH2:25][OH:26])=[CH:9]2)=[N:6][CH:7]=1
Reactant: O=C(c1ccc2c(CO)cn(-c3ncc(Br)cn3)c2c1)N1CCOCC1
Reactant: OB(O)c1ccccc1F
Product: O=C(c1ccc2c(CO)cn(-c3ncc(-c4ccccc4F)cn3)c2c1)N1CCOCC1

Procedure:
Prepared from 27c) (400 mg, 0.959 mmol, 1.0 eq) and (2-fluorophenyl)boronic acid (133 mg, 0.959 mmol, 1.0 eq) according to general procedure 1. White solid. Yield: 230 mg (55% of theory). Melting range: 190-194° C. HPLC (method 1): Rt=9.69 min. Mass spectroscopy: m/z: [M+H]+=433.0
Reaction SMILES: [F:1][C:2]([F:27])([F:26])[CH2:3][N:4]1[C:8]([C:9]2[N:10]=[C:11]3[C:17]4[CH:18]=[C:19]([C:22](O)=[O:23])[CH:20]=[CH:21][C:16]=4[O:15][CH2:14][CH2:13][N:12]3[CH:25]=2)=[N:7][CH:6]=[N:5]1.[N:28]1([CH2:34][CH2:35][OH:36])[CH2:33][CH2:32][NH:31][CH2:30][CH2:29]1>>[OH:36][CH2:35][CH2:34][N:28]1[CH2:33][CH2:32][N:31]([C:22]([C:19]2[CH:20]=[CH:21][C:16]3[O:15][CH2:14][CH2:13][N:12]4[CH:25]=[C:9]([C:8]5[N:4]([CH2:3][C:2]([F:26])([F:1])[F:27])[N:5]=[CH:6][N:7]=5)[N:10]=[C:11]4[C:17]=3[CH:18]=2)=[O:23])[CH2:30][CH2:29]1
Reactant: OCCN1CCNCC1
Reactant: O=C(O)c1ccc2c(c1)-c1nc(-c3ncnn3CC(F)(F)F)cn1CCO2
Product: O=C(c1ccc2c(c1)-c1nc(-c3ncnn3CC(F)(F)F)cn1CCO2)N1CCN(CCO)CC1

Procedure:
Following the procedure for 109, 2-(1-(2,2,2-trifluoroethyl)-1H-1,2,4-triazol-5-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepine-10-carboxylic acid and 2-(piperazin-1-yl)ethanol gave 122. MS: (ESI+)=492.2. 1H NMR (400 MHz, DMSO) δ 8.39 (d, J=2.0 Hz, 1H), 8.10 (d, J=5.4 Hz, 2H), 7.38 (dd, J=8.4, 2.1 Hz, 1H), 7.13 (d, J=8.4 Hz, 1H), 5.89 (q, J=8.7 Hz, 2H), 4.57 (s, 4H), 4.40 (t, J=5.4 Hz, 1H), 4.06 (q, J=5.3 Hz, 1H), 3.51 (dd, J=11.6, 6.0 Hz, 4H), 3.17 (d, J=5.2 Hz, 1H), 2.46-2.39 (m, 5H)
Reaction SMILES: [O:1]1[CH2:6][CH2:5][CH2:4][O:3][CH:2]1[C:7]1[CH:12]=[CH:11][C:10]([C:13]2[S:14][C:15]3[C:20]([N:21]=2)=[CH:19][CH:18]=[C:17]([Sn](C)(C)C)[N:16]=3)=[C:9]([F:26])[CH:8]=1.[O:27]1[CH2:32][CH2:31][CH:30]([C:33](Cl)=[O:34])[CH2:29][CH2:28]1>C1(C)C=CC=CC=1>[O:1]1[CH2:6][CH2:5][CH2:4][O:3][CH:2]1[C:7]1[CH:12]=[CH:11][C:10]([C:13]2[S:14][C:15]3[C:20]([N:21]=2)=[CH:19][CH:18]=[C:17]([C:33]([CH:30]2[CH2:31][CH2:32][O:27][CH2:28][CH2:29]2)=[O:34])[N:16]=3)=[C:9]([F:26])[CH:8]=1
Reactant: O=C(Cl)C1CCOCC1
Reactant: C[Sn](C)(C)c1ccc2nc(-c3ccc(C4OCCCO4)cc3F)sc2n1
Solvent: Cc1ccccc1
Product: O=C(c1ccc2nc(-c3ccc(C4OCCCO4)cc3F)sc2n1)C1CCOCC1
Temperature: 80 °C

Procedure:
A solution of 2-(4-(1,3-dioxan-2-yl)-2-fluorophenyl)-5-(trimethylstannyl)-thiazolo[5,4-b]pyridine (1.00 g, 2.087 mmol) and tetrahydro-2H-pyran-4-carbonyl chloride (0.434 mL, 2.92 mmol) in 15 mL toluene was sealed and heated to 80° C. After 4 h a precipitate formed. The reaction mixture was cooled and filtered, rinsing with Et2O, and the solid was dried in vacuo to give (2-(4-(1,3-dioxan-2-yl)-2-fluorophenyl)-thiazolo[5,4-b]pyridine-5-yl)(tetrahydro-2H-pyran-4-yl)methanone as a white solid. MS (ESI) m/z: Calculated: 428.1; Observed: 428.8 (M++1).
Reaction SMILES: Cl[C:2]1[C:7]([C:8]#N)=[CH:6][C:5]([O:10][CH3:11])=[CH:4][N:3]=1.[C:12]([O:16][CH3:17])(=[O:15])[CH2:13][SH:14].CC([O-])(C)C.[K+].C1COCC1.C[N:30](C=O)C>>[NH2:30][C:13]1([C:12]([O:16][CH3:17])=[O:15])[S:14][C:2]2=[N:3][CH:4]=[C:5]([O:10][CH3:11])[CH:6]=[C:7]2[CH2:8]1
Reactant: COC(=O)CS
Reactant: CN(C)C=O
Reactant: COc1cnc(Cl)c(C#N)c1
Agent: CC(C)(C)[O-]
Agent: [K+]
Solvent: C1CCOC1
Product: COC(=O)C1(N)Cc2cc(OC)cnc2S1
Reaction time: 0.33 h
Yield: 71 %

Procedure:
To 2-chloro-3-cyano-5-methoxypyridine (0.53 g) and methyl thioglycolate (280 μL) in DMF (10 mL) at 5° C. was added a 1.0M solution of KOtBu/THF (3.8 mL). The reaction was stirred 20 min at 5°, then 2 h at RT, quenched in sat'd NH4Cl, the solid precipitate was collected, washed with water and dried to give 0.53 g (71%) of the title compound.
Reaction SMILES: [Br-].[O:2]([CH2:9][CH2:10][CH2:11][CH2:12][CH2:13][CH2:14][P+](C1C=CC=CC=1)(C1C=CC=CC=1)C1C=CC=CC=1)[C:3]1[CH:8]=[CH:7][CH:6]=[CH:5][CH:4]=1.C[Si]([N-][Si](C)(C)C)(C)C.[K+].[CH3:44][O:45][C:46]1[CH:53]=[CH:52][CH:51]=[CH:50][C:47]=1[CH:48]=O>C1COCC1>[CH3:44][O:45][C:46]1[CH:53]=[CH:52][CH:51]=[CH:50][C:47]=1[CH:48]=[CH:14][CH2:13][CH2:12][CH2:11][CH2:10][CH2:9][O:2][C:3]1[CH:4]=[CH:5][CH:6]=[CH:7][CH:8]=1
Reactant: COc1ccccc1C=O
Reactant: c1ccc(OCCCCCC[P+](c2ccccc2)(c2ccccc2)c2ccccc2)cc1
Agent: C[Si](C)(C)[N-][Si](C)(C)C
Agent: [Br-]
Agent: [K+]
Solvent: C1CCOC1
Product: COc1ccccc1C=CCCCCCOc1ccccc1

Procedure:
1-(2-Methoxyphenyl)-7-phenoxy-1-heptene (7.3) was synthesized as described in 7.1 using 6.1 (2.0 g, 3.85 mmol), dry THF (30 mL), potassium bis(trimethylsilyl)amide (0.77 g, 3.85 mmol), and 2-methoxybenzaldehyde (0.20 g, 1.47 mmol). The title compound (7.3) was isolated as a colorless liquid after purification by flash column chromatography (0.396 g, 91% yield, predominantly cis, cis:trans=93:7).
Reaction SMILES: O.[OH-].[Li+].[Cl:4][C:5]1[CH:27]=[C:26]([C:28]([NH:30][C@@H:31]([C:33]2[C:42]3[C:37](=[CH:38][CH:39]=[CH:40][CH:41]=3)[CH:36]=[CH:35][CH:34]=2)[CH3:32])=[O:29])[CH:25]=[C:24]([Cl:43])[C:6]=1[C:7]([NH:9][C@H:10]([C:20]([O:22]C)=[O:21])[CH2:11][NH:12][C:13]([C:15]1[S:16][CH:17]=[CH:18][CH:19]=1)=[O:14])=[O:8]>O.O1CCCC1.CO>[Cl:4][C:5]1[CH:27]=[C:26]([C:28]([NH:30][C@@H:31]([C:33]2[C:42]3[C:37](=[CH:38][CH:39]=[CH:40][CH:41]=3)[CH:36]=[CH:35][CH:34]=2)[CH3:32])=[O:29])[CH:25]=[C:24]([Cl:43])[C:6]=1[C:7]([NH:9][C@H:10]([C:20]([OH:22])=[O:21])[CH2:11][NH:12][C:13]([C:15]1[S:16][CH:17]=[CH:18][CH:19]=1)=[O:14])=[O:8]
Reactant: COC(=O)[C@H](CNC(=O)c1cccs1)NC(=O)c1c(Cl)cc(C(=O)N[C@H](C)c2cccc3ccccc23)cc1Cl
Agent: [Li+]
Agent: [OH-]
Solvent: CO
Solvent: O
Solvent: C1CCOC1
Product: C[C@@H](NC(=O)c1cc(Cl)c(C(=O)N[C@@H](CNC(=O)c2cccs2)C(=O)O)c(Cl)c1)c1cccc2ccccc12
Temperature: 25 °C
Reaction time: 8 h
Yield: 67 %

Procedure:
A solution of lithium hydroxide monohydrate (100 mg, 2.38 mmol) in water (5 mL) was added to a solution of N-[2,6-dichloro-4-[[[(1R)-1-(1-naphthalenyl)ethyl]amino]carbonyl]benzoyl]-3-[(thiophene-2-carbonyl)amino]-L-alanine, methyl ester (260 mg, 0.434 mmol) in tetrahydrofuran (15 mL) and methanol (5 mL), and the resulting mixture was stirred at room temperature overnight. The solution was evaporated, and the residue was partitioned between ethyl acetate (40 mL) and 1 M HCl (10 mL). The organic layer was evaporated and the residue was dissolved in methanol (˜1 mL), purified by HPLC, and lyophilized to give N-[2,6-dichloro-4-[[[(1R)-1-(1-naphthalenyl)ethyl]amino]carbonyl]benzoyl]-3-[(thiophene-2-carbonyl)amino]-L-alanine (170 mg, 67%).
Reaction SMILES: [NH2:1][C@H:2]([C:8]([N:10]1[CH2:21][CH2:20][CH2:19][C@H:11]1[C:12]([NH:14][CH2:15][C:16]([OH:18])=[O:17])=[O:13])=[O:9])[CH2:3][CH2:4][C:5](=[O:7])[NH2:6].CCN(C(C)C)C(C)C.[NH:31](C(OCC1C=CC=CC=1)=O)[C@H:32]([C:36](ON1C(=O)CCC1=O)=[O:37])[CH:33]([CH3:35])[CH3:34].[H][H]>CC#N.[Pd].O>[NH2:31][C@H:32]([C:36]([NH:1][C@H:2]([C:8]([N:10]1[CH2:21][CH2:20][CH2:19][C@H:11]1[C:12]([NH:14][CH2:15][C:16]([OH:18])=[O:17])=[O:13])=[O:9])[CH2:3][CH2:4][C:5](=[O:7])[NH2:6])=[O:37])[CH:33]([CH3:35])[CH3:34]
Reactant: NC(=O)CC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(=O)O
Reactant: CC(C)[C@H](NC(=O)OCc1ccccc1)C(=O)ON1C(=O)CCC1=O
Reactant: [H][H]
Agent: [Pd]
Solvent: CC#N
Solvent: O
Solvent: CCN(C(C)C)C(C)C
Product: CC(C)[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(=O)NCC(=O)O
Temperature: 25 °C

Procedure:
H-Gln-Pro-Gly-OH (1 eq.) was added to H2O containing DIPEA (2.00 eq.). The slurry was stirred at 25° C. until a clear solution was observed and was then cooled to 0° C. Z-Val-OSu (1.1 eq.) was dissolved in MeCN at 25° C. until a clear solution was obtained and was then cooled to 0° C. The Z-Val-OSu solution was added to the solution of H-Gln-Pro-Gly-OH in such way that the temperature did not rise above 5° C. Then the mixture was stirred for at least 2 h. The peptide solution was concentrated and then diluted with a KHSO4 solution and stirred for a few minutes. This aqueous solution was washed twice with a mixture of IPE and AcOEt. The organic phases were discarded and the aqueous phase was extracted 2 times with a 20% i-BuOH in DCM solution. The organic phases were collected and concentrated under reduced pressure until water content in the evaporates was ≦1% weight. The solution of the protected tetrapetide was then precipitated in IPE at 25° C. Z-Val-Gln-Pro-Gly-OH (SEQ ID NO 16) was collected by filtration washed with IPE and dried under vacuum until IPE content in the peptide was <5% weight. The protected fragment was dissolved in methanol at 30° C. and Pd/C (0.02 eq.) was added to the peptide solution. The solution was stirred at 20° C. followed by the introduction of hydrogen under pressure (0.3 bar). After stirring for 3 hours the solution was filtered to remove the catalyst which was washed with methanol. After evaporation, the free tetrapeptide was then precipitated by transferring the solution into MeCN at 10° C. The solid was filtered and washed with MeCN. After drying under vacuum, 60% of H-Val-Gln-Pro-Gly-OH (SEQ ID NO 4) was recovered.
Reaction SMILES: [NH2:1][C:2]1[CH:11]=[C:10]([N:12]2[CH2:17][CH2:16][N:15]([C:18]([NH:20][CH:21]3[CH2:26][CH2:25][CH2:24][CH:23]([C:27](O)=[O:28])[CH2:22]3)=[O:19])[CH2:14][CH2:13]2)[C:9]2[C:4](=[CH:5][C:6]([Cl:30])=[CH:7][CH:8]=2)[N:3]=1.[CH3:31][N:32](C(ON1N=NC2C=CC=NC1=2)=[N+](C)C)C.F[P-](F)(F)(F)(F)F.CN.C(N(C(C)C)CC)(C)C>CN(C=O)C.C(OCC)(=O)C>[NH2:1][C:2]1[CH:11]=[C:10]([N:12]2[CH2:17][CH2:16][N:15]([C:18]([NH:20][CH:21]3[CH2:26][CH2:25][CH2:24][CH:23]([C:27]([NH:32][CH3:31])=[O:28])[CH2:22]3)=[O:19])[CH2:14][CH2:13]2)[C:9]2[C:4](=[CH:5][C:6]([Cl:30])=[CH:7][CH:8]=2)[N:3]=1
Reactant: Nc1cc(N2CCN(C(=O)NC3CCCC(C(=O)O)C3)CC2)c2ccc(Cl)cc2n1
Reactant: CN(C)C(On1nnc2cccnc21)=[N+](C)C
Agent: CN
Agent: F[P-](F)(F)(F)(F)F
Solvent: CCN(C(C)C)C(C)C
Solvent: CN(C)C=O
Solvent: CCOC(C)=O
Product: CNC(=O)C1CCCC(NC(=O)N2CCN(c3cc(N)nc4cc(Cl)ccc34)CC2)C1
Temperature: 25 °C
Reaction time: 3 h

Procedure:
The mixture of 3-[[[4-(2-amino-7-chloro-4-quinolinyl)-1-piperazinyl]carbonyl]amino]cyclohexanecarboxylic acid (30 mg, 0.069 mmol), HATU (100 mg), methylamine (1.0M in THF, 3 mL) and diisopropylethylamine (0.5 mL) in DMF (3 mL) was stirred at rt for 3 h. The reaction mixture was diluted with ethyl acetate (100 mL), washed with sat. NaHCO3 solution and water, and dried over Na2SO4. Concentration in vacuo followed by flash chromatography afforded the title product. LC-MS: 445 (M++1). 1H NMR (CD3OD) δ 1.10–1.35 (m, 4H), 1.65–1.90 (m, 4H), 2.20 (m, 1H), 2.65 (s, 3H), 3.08 (m, 4H), 3.60 (m, 5H), 5.80 (d, 1H), 6.15 (s, 1H), 7.10 (d, 2H), 7.45 (s, 1H), 7.65 (a, 1H).
Reaction SMILES: [CH3:1][N:2]1[C:7](=[S:8])[NH:6][C:5](=[O:9])[C:4](=[O:10])[NH:3]1.FC(F)(F)S(O[CH2:17][C@@H:18]1[O:22][CH2:21][C@@H:20]([S:23][C:24](=[O:26])[CH3:25])[CH2:19]1)(=O)=O.CCN(C(C)C)C(C)C>O1CCCC1.CN(C)C=O>[CH3:1][N:2]1[CH:7]([S:8][CH2:17][C@@H:18]2[O:22][CH2:21][C@@H:20]([S:23][C:24](=[O:26])[CH3:25])[CH2:19]2)[NH:6][C:5](=[O:9])[C:4](=[O:10])[NH:3]1
Reactant: Cn1[nH]c(=O)c(=O)[nH]c1=S
Reactant: CC(=O)S[C@@H]1CO[C@@H](COS(=O)(=O)C(F)(F)F)C1
Solvent: C1CCOC1
Solvent: CN(C)C=O
Solvent: CCN(C(C)C)C(C)C
Product: CC(=O)S[C@@H]1CO[C@@H](CSC2NC(=O)C(=O)NN2C)C1
Yield: 32.3 %

Procedure:
The title compound is prepared by the procedure of Example 23 using 0.551 g of product from Example 83 in 1 ml of tetrahydrofuran, 1.07 g of product from Example 22 in 1 ml of tetrahydrofuran, 3 ml of dimethylformamide, and 605 microliter of Hunig's base to give 0.357 g of the desired product.
Reaction SMILES: [Br:1][C:2]1[CH:7]=[CH:6][C:5]([C:8](=[O:12])[CH:9]([CH3:11])[CH3:10])=[CH:4][CH:3]=1.[CH2:13](O)[CH2:14][CH2:15][OH:16]>>[Br:1][C:2]1[CH:3]=[CH:4][C:5]([C:8]2([CH:9]([CH3:10])[CH3:11])[O:16][CH2:15][CH2:14][CH2:13][O:12]2)=[CH:6][CH:7]=1
Reactant: OCCCO
Reactant: CC(C)C(=O)c1ccc(Br)cc1
Product: CC(C)C1(c2ccc(Br)cc2)OCCCO1
Yield: 42 %

Procedure:
Using an analogous procedure to that described in Example 62, part (i), but starting from 1-bromo-4-(2-methylpropanoyl)benzene and 1,3-propanediol there was thus obtained (in 42% yield) 2-(4-bromophenyl)-2-isopropyl-1,3-dioxane (4.25g), b.p. 116° C. at 0.4 mm Hg; 1H NMR (acid free CDCl3): 0.8 (d, 3 H), 1.15-1.25 (m, 1 H), 1.85 (m, 2 H), 2.0-2.1 (m, 1 H), 3.68 (d of t, 2 H), 3.75-3.85 (m, 2 H) 7.15-7.25 (m, 2 H), 7.45-7.55 (m, 2 H): mass spectrum (CI+) 285 (M+H)+.
Reaction SMILES: Cl.[CH3:2][N:3]1[C:15]2[CH2:14][NH:13][CH:12]([C:16]([O:18][CH3:19])=[O:17])[CH2:11][C:10]=2[C:9]2[C:4]1=[CH:5][CH:6]=[CH:7][CH:8]=2.C(N(CC)CC)C.[C:27](=[S:29])=[S:28].[CH3:30]I>CO>[CH3:2][N:3]1[C:15]2[CH2:14][N:13]([C:27]([S:29][CH3:30])=[S:28])[CH:12]([C:16]([O:18][CH3:19])=[O:17])[CH2:11][C:10]=2[C:9]2[C:4]1=[CH:5][CH:6]=[CH:7][CH:8]=2
Reactant: CI
Reactant: S=C=S
Reactant: COC(=O)C1Cc2c(n(C)c3ccccc23)CN1
Agent: Cl
Solvent: CCN(CC)CC
Solvent: CO
Product: COC(=O)C1Cc2c(n(C)c3ccccc23)CN1C(=S)SC
Yield: 80.5 %

Procedure:
In the same manner as described in Example 53, methyl (3RS)-9-methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylate hydrochloride (5.61 g), triethylamine (4.86 g), carbon disulfide (1.83 g), methanol (200 ml) and methyl iodide (3.40 g) are reacted and treated. The product is recrystallized from methanol to give the title compound (5.38 g, 80.5%) as colorless needles, m.p. 170°-171° C.
Reaction SMILES: [CH:1]1[C:10]2[C:5](=[CH:6][CH:7]=[CH:8][CH:9]=2)[CH:4]=[CH:3][C:2]=1[CH:11]=O.[N+:13]([CH3:16])([O-:15])=[O:14].C([O-])(=O)C.[NH4+]>C(O)(=O)C>[N+:13]([CH:16]=[CH:11][C:2]1[CH:3]=[CH:4][C:5]2[C:10](=[CH:9][CH:8]=[CH:7][CH:6]=2)[CH:1]=1)([O-:15])=[O:14]
Reactant: C[N+](=O)[O-]
Reactant: O=Cc1ccc2ccccc2c1
Agent: CC(=O)[O-]
Agent: [NH4+]
Solvent: CC(=O)O
Product: O=[N+]([O-])C=Cc1ccc2ccccc2c1
Temperature: 0 °C
Yield: 94.1 %

Procedure:
A mixture of 2-naphthaldehyde (12 g), nitromethane (13.8 g) and ammonium acetate (5 g) in acetic acid (50 ml) was refluxed for 21/2hours. The brown mixture was poured into ice-water (100 ml) from which a yellow solid separated. Filtration and drying in vacuo afforded the title compound (14.4 g).
Reaction SMILES: [CH3:1][C:2]([CH3:29])([O:4][C:5]([NH:7][C@H:8]([C:17]([NH:19][C@@H:20]([C:25]([O:27]C)=[O:26])[CH2:21][CH2:22][S:23][CH3:24])=[O:18])[CH2:9][C:10]1[CH:15]=[CH:14][C:13]([OH:16])=[CH:12][CH:11]=1)=[O:6])[CH3:3].[OH-].[K+].OS([O-])(=O)=O.[K+].C(Cl)Cl>O1CCCC1.O>[CH3:3][C:2]([CH3:29])([O:4][C:5]([NH:7][C@H:8]([C:17]([NH:19][C@@H:20]([C:25]([OH:27])=[O:26])[CH2:21][CH2:22][S:23][CH3:24])=[O:18])[CH2:9][C:10]1[CH:15]=[CH:14][C:13]([OH:16])=[CH:12][CH:11]=1)=[O:6])[CH3:1]
Reactant: COC(=O)[C@@H](CCSC)NC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)OC(C)(C)C
Agent: O=S(=O)([O-])O
Agent: [K+]
Agent: [OH-]
Solvent: ClCCl
Solvent: C1CCOC1
Solvent: O
Product: CSCC[C@@H](NC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)OC(C)(C)C)C(=O)O
Reaction time: 4 h
Yield: 103.4 %

Procedure:
The title product of Example 1 (25.0 g, 58.6 mmol) dissolved in 150 ml of tetrahydrofuran (THF) was combined with a solution of potassium hydroxide (KOH, 7.7 g, 0.12 mmol) dissolved in 375 ml of water. After this reaction was stirred for 4 h, it was poured into a mixture of 2.5N KHSO4 (400 ml) and CH2Cl2 (500 ml). The organic layer was separated and the aqueous (pH2) was extracted 3×100 ml of CH2Cl2. The combined organics were washed with 150 ml of brine, dried over Na2SO4, and stripped of all solvent under reduced pressure. The residue, after in vacuo drying, yielded 25 g of the title product as a white powder. This material was used without further purification.
Reaction SMILES: P(OC[C@H]1[O:19][C@@H:18](N2C=CC(N)=NC2=O)[CH2:17][C@@H:16]1[OH:28])(OP(OP(O)(O)=O)(O)=O)(=O)[OH:2].P([O:41][CH2:42][C@H]1[O:47][C@@H:46](N2C3N=CN=C(N)C=3N=C2)[CH2:45][C@@H]1O)(OP(OP(O)(O)=O)(O)=O)(=O)O.C1N=C(N)C2N=CN([C@@H]3O[C@H](COP(OP(OP(O)(O)=O)(O)=O)(O)=O)[C@@H](C=O)[C@H]3C=O)C=2[N:60]=1>>[CH2:42]([OH:41])[C:17]([NH2:60])([CH2:16][OH:28])[CH2:18][OH:19].[C:46]([O-:47])(=[O:2])[CH3:45]
Reactant: Nc1ncnc2c1ncn2[C@H]1C[C@H](O)[C@@H](COP(=O)(O)OP(=O)(O)OP(=O)(O)O)O1
Reactant: Nc1ccn([C@H]2C[C@H](O)[C@@H](COP(=O)(O)OP(=O)(O)OP(=O)(O)O)O2)c(=O)n1
Reactant: Nc1ncnc2c1ncn2[C@@H]1O[C@H](COP(=O)(O)OP(=O)(O)OP(=O)(O)O)[C@@H](C=O)[C@H]1C=O
Product: CC(=O)[O-]
Product: NC(CO)(CO)CO
Temperature: 65 °C
Reaction time: 1 h

Procedure:
A second treatment of the DNA with the Klenow fragment of E. coli DNA polymerase I was conducted as described above except that 0.05 mM deoxycytidine triphosphate was substituted for the 0.05 mM deoxyadenosine triphosphate of the previous reaction. This produced a fragment with the sequence ##STR19## near the beginning of the rennin gene. After phenol extraction, ether extraction, and ethanol precipitation, the DNA was redissolved in water and treated with S1 nuclease (Boehringer/Mannheim, 100 units) for 30 minutes at room temperature i a 50 μl reaction containing Buffer S. This enzyme removed the 5' single-stranded DNA from the fragment leaving the sequence ##STR20## Thus, the beginning of the rennin sequence is at the 5' end of the DNA fragment. A synthetic oligonucleotide containing a ClaI restriction site and ending with the nucleotides ATG (i.e. CATCGATG, Collaborative Research, Inc., 5 μg) was kinased with Y32 -P-ATP using T4 polynucleotide kinase ((P-L Biochemicals, 3 units) for 30 minutes at 37° C. in a 25 μl reaction containing Buffer Y. This kinased linker (about 200 pmoles was ligated to the treated DNA fragment (about 5 pmoles) by incubation with T4DNA Ligase (N.E. Biolabs, 900 units) at 15° C. overnight in Buffer L. The reaction was terminated by heating at 65° C. for 5 minutes. Four microliters of 10x ClaI buffer (1x=10 mM Tris·HCl pH 8, 10 mM MgCl2), and 10 μl of restriction endonuclease ClaI (Boehringer/Mannheim, 27 units) were added. The resulting mixture wax incubated at 37° C. for one hour. Four microliters were removed for analysis on a polyacrylamide gel, followed by the addition of 1 μl of 10x ClaI buffer, 10 μl of ClaI enzyme, and 3 μl water. This mixture was incubated for an additional hour. The treated DNA containing the desired rennin sequences was purified by separation in a 2% agarose gel containing 40 mM Tris·acetate buffer pH 8.3. The DNA was visualized by long wave ultraviolet irradiation and removed from the gel by the freeze-thaw method described above. This fragment was then ready for insertion into the appropriate vector.
Reaction SMILES: [Cl:1][C:2]1[CH:7]=[CH:6][CH:5]=[CH:4][C:3]=1[CH:8]=[C:9]([C:11]1[CH:16]=[CH:15][C:14]([F:17])=[CH:13][C:12]=1[F:18])[CH3:10].[Br:19]N1C(=O)CCC1=O.C(OOC(=O)C1C=CC=CC=1)(=O)C1C=CC=CC=1>CCOC(C)=O>[Br:19][CH2:10][C:9]([C:11]1[CH:16]=[CH:15][C:14]([F:17])=[CH:13][C:12]=1[F:18])=[CH:8][C:3]1[CH:4]=[CH:5][CH:6]=[CH:7][C:2]=1[Cl:1]
Reactant: CC(=Cc1ccccc1Cl)c1ccc(F)cc1F
Reactant: O=C1CCC(=O)N1Br
Agent: O=C(OOC(=O)c1ccccc1)c1ccccc1
Solvent: CCOC(C)=O
Product: Fc1ccc(C(=Cc2ccccc2Cl)CBr)c(F)c1
Temperature: 25 °C
Yield: 85 %

Procedure:
A stirred solution of 1-[1-(2-chlorophenyl)prop-1-en-2-yl]-2,4-difluorobenzene (680 mg, 2.56 mmol), N-bromosuccinimide (823 mg, 4.62 mmol) and benzoyl peroxide (30 mg) in AcOEt (15 ml) was heated under reflux for 3 h. After cooling to room temperature, the reaction mixture was diluted with AcOEt and washed with water, and the combined organic phases were dried and concentrated to dryness under reduced pressure. Purification by chromatography on silica gel gave 1-[3-bromo-1-(2-chlorophenyl)prop-1-en-2-yl]-2,4-difluorobenzene as a 52/48 mixture of diastereoisomers [colourless oil, 792 mg, yield 85%; GC/MS: m/z=344 (M+); HPLC/MS: log P(HCOOH)=4.80 (major diastereoisomer), 5.25 (minor diastereoisomer)].
Reaction SMILES: COC1C=CC(CN[CH:9]=[N:10][NH:11][C:12]([O:14][CH3:15])=[O:13])=CC=1.Br[CH2:19][C:20]([C:22]1[CH:23]=[N:24][N:25]([CH3:28])[C:26]=1[Br:27])=[O:21].C(=O)([O-])[O-:30].[Na+].[Na+].C(N(CC)C(C)C)(C)C>C(#N)C.O>[Br:27][C:26]1[N:25]([CH3:28])[N:24]=[CH:23][C:22]=1[C:20](=[O:21])[CH2:19][N:10]([CH:9]=[O:30])[NH:11][C:12]([O:14][CH3:15])=[O:13]
Reactant: O=C([O-])[O-]
Reactant: COC(=O)NN=CNCc1ccc(OC)cc1
Reactant: Cn1ncc(C(=O)CBr)c1Br
Agent: [Na+]
Solvent: CCN(C(C)C)C(C)C
Solvent: O
Solvent: CC#N
Product: COC(=O)NN(C=O)CC(=O)c1cnn(C)c1Br
Reaction time: 0.17 h
Yield: 27.2 %

Procedure:
Methyl 2-{[(4-methoxybenzyl)amino]methylene}hydrazinecarboxylate (3.28 g, 13.8 mmol), 2-bromo-1-(5-bromo-1-methyl-1H-pyrazol-4-yl)ethanone (3.90 g, 13.8 mmol) and sodium carbonate (1.16 g, 13.8 mmol) were combined in a mixture of N,N-diisopropylethylamine (99.5%, 2.30 mL, 13.8 mmol) and acetonitrile (30 mL). The reaction was heated at 80° C. for 18 hours, at which time water was added, and heated was continued for 10 minutes. After removal of solvent in vacuo, the aqueous residue was partitioned between water and ethyl acetate (100 mL). The aqueous layer was extracted with ethyl acetate (4×100 mL), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. Purification using silica gel chromatography (Gradient: 0% to 100% ethyl acetate in heptane) provided the product (1.20 g), contaminated with some impurities. This material was taken directly to the following step. LCMS m/z 319.0 (M+1). 1H NMR (400 MHz, CD3OD) δ, product peaks: 3.74 (s, 3H), 3.92 (s, 3H), 4.82 (br s, 2H), 8.14 (s, 1H), 8.24 (s, 1H).
Reaction SMILES: [CH:1]1([C:7]([S:9]C)=S)[CH2:6][CH2:5][CH2:4][CH2:3][CH2:2]1.[O:11]=[C:12]1[N:17]2[CH2:18][CH2:19][CH2:20][C:21]3[CH:26]=[CH:25][CH:24]=[CH:23][C:22]=3[CH:16]2[CH2:15][NH:14][CH2:13]1.O>CN(C)C=O>[CH:1]1([C:7]([N:14]2[CH2:13][C:12](=[O:11])[N:17]3[CH2:18][CH2:19][CH2:20][C:21]4[CH:26]=[CH:25][CH:24]=[CH:23][C:22]=4[CH:16]3[CH2:15]2)=[S:9])[CH2:2][CH2:3][CH2:4][CH2:5][CH2:6]1
Reactant: O=C1CNCC2c3ccccc3CCCN12
Reactant: CSC(=S)C1CCCCC1
Solvent: O
Solvent: CN(C)C=O
Product: O=C1CN(C(=S)C2CCCCC2)CC2c3ccccc3CCCN12

Procedure:
Methyl cyclohexanecarbodithioate (0.24 g) was added to a solution of 4-oxo-1,2,3,4,6,7,8,12b-octahydropyrazino[2,1-a][2]benzazepine (0.3 g) in dimethylformamide (5 ml). The mixture was refluxed for 4 h, cooled and poured into water. This was extracted with diethyl ether; the ether solution was dried (MgSO4) and evaporated. The residue was purified by column chromatography (SiO2, diethyl ether).